Product and process-related impurities impact biopharmaceutical development at every stage from discovery to development to manufacturing. Detecting, identifying and characterizing these variants and impurities are of high regulatory concern, due to the
potential correlation between aggregation and increased immunogenicity of the biotherapeutics.
Among the impurities, aggregates such as subvisible and submicron particles, and host cell proteins are of particular challenge. Responding to popular demand, CHI is pleased to bring you a new conference on Characterizing Aggregates & Impurities as
part of the Biotherapeutics Analytical Summit, to bring together best practices and new insights in the identification, monitoring and characterization of these impurities and contaminants.
MONDAY, MARCH 12, 2018
7:30 am Registration and Morning Coffee
8:30 Chairperson’s Opening Remarks
Mark T. Fisher, PhD, Professor, Biochemistry and Molecular Biology, University of Kansas Medical Center
8:40 Regulatory Address: Regulatory Perspectives on Analytical Method Validation and Transfer for Biopharmaceutical Products
Ramesh Potla, PhD, Product Quality Team Leader, CDER, FDA
A risk-based approach should be used to determine the totality of evidence needed to demonstrate a successful method transfer. This presentation will focus on our current expectations about the validation and any subsequent site transfer of validated
analytical methods for biotechnology products. Case studies will be presented with the goal of sharing lessons learned during the regulatory review of non-compendial analytical method validation and transfer for licensed therapeutic proteins.
9:20 MAM Method Development and Qualification
Richard Rogers, PhD, Scientist 4, Just Biotherapeutics
We have developed and implemented a mass spectrometry based multi-attribute method (MAM) that monitors known CQAs but also can identify new CQAs on the biotherapeutics. This method has been successfully used in the process development lab. Our goal
is to leverage the MAM for release of biotherapeutics from the quality lab. Method qualification and specification limits for the MAM will be discussed in this presentation.
9:50 The Next Frontier in Subvisible Particle Analysis: New Tools and Potential Opportunities
Danny K. Chou, PharmD, PhD, President, Compassion Biosolution, LLC
In the past decade, we have witnessed the arrival of a large number of analytical technologies that are useful for characterizing sub-visible particles in protein therapeutics. Even with the diverse tools that are available today, there are still
important gaps that have not been filled but yet have a significant role in our ability to fully analyze particles for either product characterization or development purpose. The goal of this presentation is to highlight some of these gaps and
share the potential opportunities that may be captured by new tools that are on the horizon.
10:20 Networking Coffee Break
10:50 New Developments in the Characterization of Fibril Aggregates in Peptide Therapeutics: From Aggregation Kinetics to Single Nanoparticle Detection Methods
Jingtao Zhang, PhD, Principal Scientist, Pharmaceutical Sciences, Merck & Co.
The formation of irreversible aggregates such as fibrils, has proven to be a key challenge in developing synthetic peptide therapeutics. In this presentation, we will discuss the investigation on the aggregation kinetics of a fibril-prone peptide,
the projection of physical stability shelf-life, and the development of highly sensitive characterization methods for fibrils. In particular, the application of highly sensitive submicron detection tools such as Archimedes and flow cytometry instrument
to investigate the fibril behavior in lag-phase will be discussed.
11:20 CE Methods on Quantification of Adeno-Associated Virus (AAV) Capsid Purity
Wei-Chiang Chen, PhD, Scientist I, Analytical Development, Biogen
Recombinant adeno-associated virus (AAV) was demonstrated as a promising platform in human gene therapy. AAV capsids are comprised of three viral proteins, VP1, VP2, and VP3, and their theoretical ratio is 1:1:10. Capillary electrophoresis (CE)
SDS has been widely used to analyze fragments or impurities in biologics. In this study, we demonstrate successful development of AAV capsid purity assay on multiple CE platforms which are commonly used in industry. The results from different
CE platforms are compared and summarized in this presentation.
11:50 Characterization of Aggregates and Impurities of Proteins Used in Diagnostic Blood Screening Assays
Jeffrey Fishpaugh, PhD, Senior Principal Research Scientist, Analytical Chemistry
Biologics used in clinical diagnostics blood screening assays occupy a unique design space. These assays and their components (biologics/proteins) fall under the purview of the FDA's CBER section, the same section that inspects and reviews biotherapeutic products. Our current biologics release testing uses classic 20th century techniques. Recent characterization work identified and quantified impurities in our blood screening biologics. Follow-up activities include analysis employing more modern methods that are used to characterize biotherapeutics and biosimilars.
12:20 pm Luncheon Presentation (Sponsorship Opportunity Available) or Enjoy Lunch on Your Own
12:50 Session Break
1:40 Chairperson’s Remarks
Linda Yi, PhD, Senior Scientist, Analytical Development, Biogen
1:45 Using Nature’s Toolbox to Capture and Detect Preaggregate Transient States
Mark T. Fisher, PhD, Professor, Biochemistry and Molecular Biology, University of Kansas Medical
We’ve developed numerous automated GroEL chaperonin-based biolayer interferometry platforms to 1) detect preaggregate states of multiple biotherapeutic samples and 2) assess ligand or solution based stabilities at ambient or physiologically
relevant temperatures. The target protein states that are captured by GroEL-biosensors can be released into ul volumes where visualization of structures potential preaggregate proteins is easily accomplished using negative stain electron
tomography and can be identified using Mass spectroscopy.
2:15 Functional, Biophysical and Structural Characterization of Engineered Antibody Variants and Quantitative Correlation between Self-Association and Particle Size in Early Stage Discovery
Sam Wu, PhD, Principal Scientist, Janssen Biotherapeutics, Janssen R&D, LLC
An early-stage developability workflow was designed to set stage gate for molecules progressing to late-stage development. Application of solubility and stability analysis in antibody selection will be presented in this study. Stability
analysis of engineered antibody variants was evaluated using Intrinsic Fluorescence Conformational StabilityTM and Colloidal stability (Tagg). B22/kD parameters for monoclonal antibodies (mAbs) in solution were determined and validated
by Small-Angle-X ray-scattering (SAXS). Quantitative correlation between Tagg and particle size for mAbs was observed in this study.
2:45 Networking Refreshment Break
3:15 Prediction of Protein Solubility for Biologics Development: From Basics to Practice
Ying Wang, PhD, Assistant Professor, Chemistry, University of North Carolina at Wilmington
One challenge in biologics development is that proteins may lose their solubility and form aggregates, crystals, viscous liquids, or gels. The protein solubility problems are frequently encountered throughout biologics development: from
early developability evaluation and formulation to manufacture and purification DOE. I will talk about the universal mechanism underlying various protein solubility issues. Then, I will show how to predict short-term and long-term
solubility problems with a simple quick experiment.
3:45 Monitoring Oxidative Damage in Therapeutic Proteins by Spectroscopic Methods
Sambit Kar, PhD, Principal Scientist & Head, Biophysics Center of Excellence, Molecular
& Analytical Development, Bristol-Myers Squibb Co.
This presentation will discuss oxidative stress on therapeutiic proteins and share strategies for monitoring degradation pathways and identifying degradation products.
4:15 Roundtable Breakout Session
Rapid Evaluation of Therapeutic Protein Preaggregate States Using Microscale Biolayer Interferometry Methodologies
Moderator: Mark T. Fisher, PhD, Biochemistry and Molecular Biology, University of Kansas Medical Center
- Detection of preaggregate states with GroEL Biosensors
- Release and evaluation of captured proteins using Electron microscopy and Mass spectroscopy
- Stability assessments using Denaturant pulse Biolayer interferometry
- Future Expansion of these methods
CQA Assessment, Control Strategies and Specification Setting
Moderator: Sambit Kar, PhD, Principal Scientist & Head, Biophysics Center of Excellence, Molecular & Analytical Development, Bristol-Myers Squibb Co.
5:15 Close of Day/Short Course Registration
6:00 Dinner Short Courses*
SC1: Particles in Biotherapeutics: Characterization & Impact
SC2: The Multi-Attribute Method (MAM) for Improving Product and Process Development
*Separate registration applies.
TUESDAY, MARCH 13, 2018
8:00 am Morning Coffee
8:30 Chairperson’s Opening Remarks
Sambit Kar, PhD, Principal Scientist & Head, Biophysics Center of Excellence, Molecular & Analytical Development, Bristol-Myers Squibb Co.
8:40 Factors Influencing Biotherapeutic Monoclonal Antibody Aggregation
Linda Yi, PhD, Senior Scientist, Analytical Development, Biogen
Aggregation has been identified as one of the major degradation pathways that may affect safety, quality and efficacy of therapeutic mAbs. Aggregates present in mAb products can be complex, varying by size, type and origin, with underline
mechanisms not always being well-understood. This presentation will provide an overview of the factors that may influence biotherapeutic mAb aggregation. A case study will follow on impact of a chemical modification catalyzed by
metals on aggregation of a few mAbs.
9:10 Characterization of Subvisible Particles: Old Challenges and Newest Improvements
Anacelia Rios Quiroz, PhD, Scientist, Group Leader Particle Lab,
Pharma Technical Development Europe-Analytics Biochemistry (PTDE-A), F. Hoffmann-La Roche Ltd.
The talk will give an overview on commercially available counting methodologies for detection of subvisible particles (SbVP). This species, ubiquitously present in protein formulations, had been in focus due to immunogenicity and quality
attributes of biotechnological products. Thus, the analytical toolbox to characterize them undergoes constant renewals and innovations. Their applicability towards the assessment of a meaningful array for particle counting characterization
will be discussed including examples of their use in the frame of immunogenicity studies.
9:40 Challenges in Characterization of Subvisible Particles in High Concentration Protein Formulations
Miguel Saggu, PhD, Scientist, Late Stage Pharmaceutical Development, Genentech, Inc.
Regulatory agencies require manufacturers of protein therapeutics to control subvisible particles in drug products to ensure the safety and efficacy of the drug as well as to demonstrate process consistency. This talk will cover case
studies of challenges in particle characterization of high concentration mAb samples.
10:10 Best Practices and Strategies for Host Cell Protein ELISAs
Alla Zilberman, Technical Marketing Manager, Cygnus Technologies
Regulatory agencies around the world expect sponsors to have a good understanding of the HCP profile of their Drug Product. Knowing that low HCP results are due to HCP content and not due to an insensitive HCP ELISA is key. Talk will
highlight current best practices and strategies to effectively demonstrate that an HCP ELISA is fit for purpose, and focus on Antibody Affinity Extraction (AAE) as a superior method for demonstrating HCP antibody coverage and reactivity
to HCPs that persist through the purification process.
10:40 Opening Coffee Break in the Exhibit Hall with Poster Viewing
11:20 Characterization of Process-Related Impurities for Next-Generation Antibody-Based Therapeutics
Jennifer Kessler, MSc, Development Associate III, Macrogenics, Inc.
Monoclonal antibodies and bispecific DART(R) molecules are being developed for a variety of indications including immuno-oncology. A risk-based approach for analytical characterization of process related impurities is required to ensure product quality of pipeline molecules during development. This presentation will discuss characterization of process-related impurities using this novel class of molecules and other antibody molecules as case studies.
11:50 New Standards for Protein Particulates
Dean Ripple, PhD, Leader, Bioprocess Measurements Group, National Institute of
Standards and Technology
12:20 pm Close of Characterizing Aggregates & Impurities